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Publication numberUS7485454 B1
Publication typeGrant
Application numberUS 09/707,852
Publication dateFeb 3, 2009
Filing dateNov 7, 2000
Priority dateMar 10, 2000
Fee statusLapsed
Also published asCA2400978A1, CN1441703A, EP1265708A1, EP1265708A4, US20040121454, US20090275115, WO2001068257A1
Publication number09707852, 707852, US 7485454 B1, US 7485454B1, US-B1-7485454, US7485454 B1, US7485454B1
InventorsAndrey Zarur Jury, Mark D. Angelino
Original AssigneeBioprocessors Corp.
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Microreactor
US 7485454 B1
Abstract
Chemical and biological reactors, including microreactors, are provided. Exemplary reactors include a plurality of reactors operable in parallel, where each reactor has a small volume and, together, the reactors produce a large volume of product. Reaction systems can include mixing chambers, heating/dispersion units, reaction chambers, and separation units. Components of the reactors can be readily formed from a variety of materials. For example, they can be etched from silicon. Components are connectable to and separable from each other to form a variety of types of reactors, and the reactors can be attachable to and separable from each other to add significant flexibility in parallel and/or series reactor operation.
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Claims(6)
1. A system for maintaining and cultivating cells in culture and obtaining a protein resulting from interaction of the cells with oxygen and/or nutrients and/or other components, comprising:
a small-scale chemical or biochemical reactor comprising a plastic substrate comprising at least one reaction unit, the reactor comprising an inlet, an outlet, and a fluid pathway connecting the inlet and the outlet, the fluid pathway comprising a chamber having a surface suitable for cell growth and a volume of less than about 1 ml, the chamber being constructed and arranged to maintain and cultivate cells in culture, the chamber further comprising an inlet fluidly connectable to a source of nutrients for the cells having a controlled pH, and an outlet for release of the protein resulting from the interaction involving the cells in the chamber;
a membrane defining at least one wall of the fluid pathway;
an enclosure positioned proximate the membrane, wherein at least one product of the interaction involving cells in the chamber passes across the membrane into the enclosure;
the reactor further comprising a pH sensor.
2. A system for maintaining and cultivating cells in culture and obtaining a protein resulting from interaction of the cells with oxygen and/or nutrients and/or other components, comprising:
a small-scale chemical or biochemical reactor comprising a plastic substrate comprising at least one reaction unit, the reactor comprising an inlet, an outlet, and a fluid pathway connecting the inlet and the outlet, the fluid pathway comprising a chamber having a surface suitable for cell growth and a volume of less than about 1 ml, the chamber being constructed and arranged to maintain and cultivate cells in culture, the chamber further comprising an inlet fluidly connectable to a source of nutrients for the cells having a controlled pH, and an outlet for release of the protein resulting from the interaction involving the cells in the chamber;
a membrane defining at least one wall of the fluid pathway;
an enclosure positioned proximate the membrane, wherein at least one product of the interaction involving cells in the chamber passes across the membrane into the enclosure;
the reactor further comprising an oxygen sensor.
3. A system as in claim 1, further comprising means for controlling the temperature of the chamber to maintain a temperature suitable for cultivating cells to generate the protein resulting from interaction of the cells with oxygen and/or nutrients and/or other components.
4. A system as in claim 1, further comprising a mixing unit fluidly connectable to the inlet of the chamber, the mixing unit including an outlet connectable to the inlet of the chamber, a plurality of inlets each in fluid communication with the outlet and a mixing chamber between plurality of inlets and of the outlet.
5. A system as in claim 4, wherein the mixing chamber is free of active mixing elements.
6. A system as in claim 2, wherein the system comprises a plurality of reaction units attachable to and separable from each other, the plurality of reaction units being constructed and arranged to operate in parallel.
Description
RELATED APPLICATION

This application claims the benefit of priority under 35 U.S.C. §119(e) of co-pending U.S. Provisional Patent Application Ser. No. 60/188,275, filed Mar. 10, 2000.

FIELD OF THE INVENTION

The present invention relates generally to chemical or biochemical microreactors, and more particularly to a microreactor for the production of the product of a chemical or biochemical reaction, including a plurality of individuated microreactors constructed to operate in parallel.

BACKGROUND OF THE INVENTION

A wide variety of reaction systems are known for the production of the product of chemical or biochemical reactions. Chemical plants involving catalysis, biochemical fermenters, pharmaceutical production plants, and a host of other systems are well-known.

Systems for housing chemical and biochemical reactions not necessarily for the production of product also are known. For example, continuous-flow systems for the detection of various analytes in bodily fluids including blood, such as oxygen, glucose, and the like are well known.

In many of these and other systems, the capacity of the system (the volume of material that the system is designed to produce, process, or analyze) is adjusted in accordance with the volume of reactant, product, or analyte desirably processed or analyzed. For example, in large-scale chemical or pharmaceutical production, reactors are generally made as large as possible to generate as large a volume of product as possible. Conversely, in many areas of clinical diagnosis, where it is desirable to obtain as much information as possible from as small a physiological sample as possible (e.g., from a tiny drop of blood), it is a goal to minimize the size of reaction chambers of sensors. Several examples of small-scale reactor systems, including those used in clinical diagnoses and other applications, follow.

U.S. Pat. No. 5,387,329 (Foos, et al.; Feb. 7, 1995) describes an extended use planar clinical sensor for sensing oxygen levels in a blood sample.

U.S. Pat. No. 5,985,119 (Zanzucchi, et al.; Nov. 16, 1999) describes small reaction cells for performing synthetic processes in a liquid distribution system. A variety of chemical reactions including catabolic, anabolic reactions, oxidation, reduction, DNA synthesis, etc. are described.

U.S. Pat. No. 5,674,742 (Northrup, et al.; Oct. 7, 1997) describes an integrated microfabricated instrument for manipulation, reaction, and detection of microliter to picoliter samples. The system purported by is suitable for biochemical reactions, particularly DNA-based reactions such as the polymerase chain reaction.

U.S. Pat. No. 5,993,750 (Ghosh, et al.; Nov. 30, 1999) describes an integrated micro-ceramic chemical plant having a unitary ceramic body formed from multiple ceramic layers in the green state which are sintered together defining a mixing chamber, passages for delivering and reacting fluids, and means for delivering mixed chemicals to exit from the device.

Biochemical processing typically involve the use of a live microorganism (cells) to produce a substance of interest. Biochemical and biomedical processing account for about 50% of the total drug, protein and raw amino-acid production worldwide. Approximately 90% of the research and development (R&D) budget in pharmaceutical industries is currently spent in biotechnology areas.

Currently bioreactors (fermentors) have several significant operational limitations. The most important being maximum reactor size which is linked to aeration properties, to nutrient distribution, and to heat transfer properties. During the progression of fermentation, the growth rate for cells accelerates, and the measures required to supply the necessary nutrients and oxygen sets physical and mechanical constraints on the vessel within which the cells are contained. Powerful and costly drives are needed to compensate for inefficient mixing and low mass-transfer rates. Additionally, as metabolism of cells accelerates, the cells generate increased heat which needs to be dissipated from the broth.

The heat transfer characteristics of the broth and the vessel (including heat exchanger) impose serious constraints on the reaction scale possible (see Table 1). While the particular heat load and power requirements are specific to the reaction, the scale of reaction generally approaches limitations as ˜10 m3 as in the case of E. coli fermentation (Table 1). The amount of heat to be dissipated becomes excessive due to limits on heat transfer coefficients of the broth and vessel. Consequently, the system of vessel and broth will rise in temperature. Unfortunately, biological compounds often have a relatively low upper limit on temperature for which to survive (<45° C. for many). Additionally, power consumption to disperse nutrients and oxygen and coolant requirements to control temperature make the process economically unfeasible (see Table 1).

TABLE 1
Oxygen- and Heat- Transfer Requirements for E. coli: Effects of Scale
OTR Volumea Pressure Power Heat Load Coolantb
(mmol/L · h) (m3) (psig) (hp) (Btu/h) (° F.)
150 1 15 5.0 84000 40
200 1 25 4.9 107000 40
300 1 35 7.1 161000 40
400 1 35 6.9 208000 40
150 10 15 50.2 884000 40
200 10 25 50.0 1078000 40
300 10 35 75.7 1621000 22
400 10 35 77.0 2096000 5
aLiquid volume
bCoolant flow is 35 gal/mm for 1-m3 vessel and 100 gal/mm for 10-m3 vessel
cCharles, M. and Wilson, J. Fermentor Design; In: Bioprocess Engineering; Lydersen, B. K., D′Elia, N. A., Nelson, K. L., Ed.; John Wiley & Sons, Inc., New York, 1994.

Aside from reactor scalability, the design of conventional fermentors has other drawbacks. Due to the batch and semi-batch nature of the process, product throughput is low. Also, the complexity and coupled nature of the reaction parameters, as well as the requirement of narrow ranges for these parameters, makes control of the system difficult. Internal to the system, heterogeneity in nutrient and oxygen distribution due to mixing dynamics creates pockets in the broth characterized by insufficient nutrients or oxygen resulting in cell death. Finally, agitation used to produce as homogeneous a solution as possible (typically involving impellar string to simultaneously mix both cells and feeds of oxygen and nutrients) causes high strains which can fracture cell membranes and cause denaturation.

While a wide variety of useful reactors for a variety of chemical and biological reactions, on a variety of size scales exist, a need exists in the art for improved reactors. In particular, there is a current need to significantly improve the design of bioreactors especially as the pharmaceutical and biomedical industries shift increasingly towards bioprocessing.

SUMMARY OF THE INVENTION

The present invention provides systems, methods, and reactors associated with small-scale chemical or biochemical reactions.

In one aspect the invention provides a chemical or biochemical reactor. The reactor includes a reaction unit including a chamber having a volume of less than one milliliter. The chamber includes an inlet connectable to a source of a chemical or biological starting material and an outlet for release of a product of a chemical or biological reaction involving the starting material. A collection chamber is connectable to the outlet of the reaction chamber. The collection chamber has a volume of greater than one liter.

In another aspect the invention involves a chemical or biochemical reactor system. The system includes a mixing chamber including a plurality of inlets connectable to a plurality of sources of chemical or biochemical reagents, and an outlet. A reaction chamber is connectable to and removable from the mixing chamber, and has a volume of less than one milliliter. The reaction chamber includes an inlet connectable to and removable from the outlet of the mixing chamber, and an outlet for release of a product of a chemical or biological reaction involving the starting material.

In another aspect the invention provides methods. One method includes carrying out a chemical or biological reaction in a plurality of reaction chambers operable in parallel, where each reaction chamber has a volume of less than one milliliter. Product of the reaction is discharged from the plurality of reaction chambers simultaneously into a collection chamber having a volume of greater than one liter.

Other advantages, novel features, and objects of the invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings, which are schematic and which are not intended to be drawn to scale. In the figures, each identical or nearly identical component that is illustrated in various figures is represented by a single numeral. For purposes of clarity, not every component is labeled in every figure, nor is every component of each embodiment of the invention shown where illustration is not necessary to allow those of ordinary skill in the art to understand the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a microbioreactor of the invention including mixing, heating/dispersion, reaction, and separation units, in expanded view;

FIG. 1A illustrates a microbioreactor of the invention including various mixing, heating/dispersion, reaction, and separation units, in expanded view;

FIG. 2 illustrates the system of FIG. 1 as assembled;

FIG. 3 illustrates the mixing unit of the system of FIG. 1;

FIG. 4 is an expanded view of the heating/dispersion unit of the system of FIG. 1;

FIG. 5 is an expanded view of the reaction chamber of the system of FIG. 1; and

FIG. 6 is an expanded view of the separation unit of the system of FIG. 1.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a chemical or biochemical reactor that can be used for a variety of very small-scale techniques. In one embodiment, a microreactor of the invention comprises a matrix of a few millimeters to a few centimeters in size containing reaction channels with dimensions on the order of hundreds of microns. Reagents of interest are allowed to flow through these microchannels, mixed, and reacted together. The products can be recovered, separated, and treated within the system. While one microreactor may be able only to hold and react a few microliters of the substances of interest, the technology allows for easy scalability and tremendous parallelization. With enhanced oxygen and nutrient distribution, a microreactor of the invention demonstrates increased performance in terms of cell viability. The microreactor geometry resembles closely the natural environment of cells whereby diffusional oxygen and nutrient transfer take place through a high surface area, thin layer interface.

With regard to throughput, an array of many microreactors can be built in parallel to generate capacity on a level exceeding that allowed by current vessels and more uniform in product quality than can be obtained in a batch method. Additionally, an advantage is obtained by maintaining production capacity at the scale of reactions typically performed in the laboratory. In general, the coupled parameters for heat and mass transfer that are determined on the lab-scale for a process do not scale linearly with volume. With conventional reactors, as the magnitude of volume is increased 1,000-1,000,000 times for production, these parameters need to be re-evaluated, often involving a large capital-investment. The use of small production volumes, although scaled in parallel, reduces the cost of current scale-up schemes.

Furthermore, the process can be implemented on a simple platform, such as an etched article for example, a silicon wafer. With the effort of semiconductor manufacturing being towards the reduction in the dimensions of channels, an opportunity to utilize excess capacity within these production facilities (with unused equipment for the larger dimensions) is provided. Mass production of these units can be carried out at very low cost and an array of many reactors, for example thousands of microreactors typically can be built for a price lower than one traditional bioreactor.

Referring now to FIG. 1, a chemical or biochemical reactor in accordance with one embodiment of the invention is illustrated schematically. The reactor of FIG. 1 is, specifically, a microbioreactor for cell cultivation. It is to be understood that this is shown by way of example only, and the invention is not to be limited to this embodiment. For example, systems of the invention can be adapted for pharmaceutical production, hazardous chemical production, or chemical remediation of warfare reagents, etc.

Microreactor 10 includes four general units. A mixing unit 12, a heating/dispersion unit 14, a reaction unit 16, and a separation unit 18. That is, in the embodiment illustrated, processes of mixing, heating, reaction, purification are implemented in series. Although not shown, pressure, temperature, pH, and oxygen sensors can be included, for example embedded within the network to monitor and provide control for the system. Due to the series format, the opportunity for several reaction units in series for multi-step chemical syntheses, for several levels of increased purification, or for micro-analysis units is provided as well.

FIG. 1 shows microreactor 10 in expanded view. As illustrated, each of units 14 and 16 (heating/dispersion and reaction units, respectively) includes at least one adjacent temperature control element 20-26 including a channel 28 through which a temperature-control fluid can be made to flow. As illustrated, temperature control units 20 and 24 are positioned above and below unit 14 and units 22 and 26 are positioned above and below unit 16. Separation unit 18 includes upper and lower extraction solvent fluid units 30 and 32, respectively, separated from unit 18 by membranes 34 and 36, respectively. FIG. 1A shows a similar microreactor having more than one reaction unit (certain elements removed for purposes of clarity).

Referring now to FIG. 2, reactor 10 is illustrated as assembled. The individual units of microreactor 10 will now be described in greater detail.

Referring now to FIG. 3, mixing unit 12 is illustrated. Mixing unit 12 is designed to provide a homogeneous mixture of starting materials or reactants to be provided to the reaction units, optionally via the heating/dispersion unit. In the specific example of the microbioreactor, mixing unit 12 is designed to provide a homogeneous broth with sufficient nutrients and oxygen, and at the required pH, for cells. Rather than combine the mixing process with simultaneous nourishment of the cells, the process is performed in a preliminary stage and then fed to the reaction stage where cells are immobilized. In this manner, the cells do not experience any shear stress due to mixing and a homogeneous mixture of feed requirements is guaranteed.

As is the case for other components of the reactor, mixing unit 12 can be manufactured using any convenient process. In preferred embodiments the unit is etched into a substrate such as silicon via known processes such as lithography. Other materials from which mixing unit 12, or other components of the systems of the invention can be fabricated, include glass, fused silica, quartz, ceramics, or suitable plastics. Silicon is preferred. The mixing unit includes a plurality of inlets 40-50 which can receive any of a variety of reactants and/or fluid carriers. Although six inlets are illustrated, essentially any number of inlets from one to tens of hundreds of inlets can be provided. Typically, less than ten inlets are needed for a given reaction. Mixing unit 12 includes an outlet 52 and, between the plurality of inlets and the outlet, a mixing chamber 54 constructed and arranged to coalesce a plurality of reactant fluids provided through the inlets. It is a feature of the embodiment illustrated that the mixing chamber is free of active mixing elements. Instead, the mixing chamber is constructed to cause turbulence in the fluids provided through the inlets thereby mixing and delivering a mixture of the fluids through the outlet without active mixing. Specifically, the mixing unit includes a plurality of obstructions 56 in the flow path that causes mixture of fluid flowing through the flow path. These obstructions can be of essentially any geometrical arrangement. As illustrated, they define small pillars about which the fluid must turbulently flow as it passes from the inlets through the mixing chamber toward the outlet. As used herein “active mixing elements” is meant to define mixing elements such as blades, stirrers, or the like which are movable relative to the reaction chamber itself, that is, movable relative to the walls defining the reaction chamber.

The volume of the mixing chamber, that is, the volume of the interior of mixing unit 12 between the inlets and the outlet, can be very small in preferred embodiments. Specifically, the mixing chamber generally has a volume of less than one liter, preferably less than about 100 microliters, and in some embodiments less than about 10 microliters. The chamber can have a volume of less than about five microliters, or even less than about one microliter.

Specifically, in the microbioreactor illustrated, six separate feed streams empty into the mixing chamber under pressure. One feed stream provides gaseous oxygen (O2) as a cell requirement. One stream, respectively, provides carbon dioxide (CO2) and nitrogen (N2) for altering pH. The remaining three channels provide the broth solution including solvent and nutrients. One of these latter streams can also be utilized to provide any additional requirements for the system such as antifoaming agents. Antifoaming agents are sometimes necessary to prevent production of foam and bubbles that can damage cells within the broth. The feed of the various streams into the chamber provides enough turbulence for mixing of the different streams. Flow within microfluidic devices is characterized by a low Reynolds number indicating the formation of lamina. While the turbulence created by the injection streams should provide sufficient mixing before the development of laminar flow, pilon-like obstructions 56 are placed in the flow path of the stream leaving the primary mixing chamber in order to enhance mixing of the lamina. By splitting a main stream into substreams followed by reunification, turbulence is introduced in the flow path, and a mechanism other than simple diffusion is used to facilitate further mixing. The length of this mixing field can be lengthened or shortened depending on the system requirements.

Referring now to FIG. 4, heating/dispersion unit 14 is shown. Unit 14 can be formed as described above with respect to other units of the invention. Unit 14 includes an inlet 60 in fluid communication with a plurality of outlets 62 in embodiments where dispersion as described below is desirable. In operation, a stream of homogeneous fluid exiting the mixing unit (feed broth in the specific microbioreactor embodiment shown) enters a dispersion matrix defined by a plurality of obstruction dividing the stream into separate flow paths directed toward the separate outlets 62. The dispersion matrix is sandwiched between two temperature control elements 20 and 24 which, as illustrated, include fluid flow channels 28 etched in a silicon article. Control unit 24 is positioned underneath unit 14, thus etched channel 28 is sealed by the bottom of unit 14. Control unit 20 is positioned atop unit 14 such that the bottom of unit 20 seals and defines the top of diffusion unit 14. A cover (not shown) can be placed a top unit 20 to seal channel 28.

Rather than for mixing, as in the previous case (FIG. 3), the splitting of the streams is to disperse the medium for its entrance into the reactive chamber in the next unit operation. In traditional reactor systems, fluid flow about a packing material containing catalysts produces the desired reaction. However, if the fluid is not evenly dispersed entering the chamber, the fluid will flow through a low resistance path through the reactor and full, active surface area will not be utilized. Dispersion in this case is to optimize reactor efficiency in the next stage.

With regard to the heating function of this unit, the platform functions as a miniaturized, traditional heat exchanger. Etched silicon platforms both above and below the central platform serve to carry a heated fluid. Cells typically require their environment to have a temperature of ˜30° C. The fluids flowing in the etched coils both above and below the broth flow channel heating the broth through the thin silicon layer. The temperature of the fluid in the upper and lower heat exchangers can be modified to ensure proper temperature for the broth. Additionally, the platform can be extended for increased heating loads.

Although a combination heating/dispersion unit is shown, unit 14 can be either a dispersion unit or a heating unit. For example, dispersion can be provided as shown, without any temperature control. Alternatively, no dispersion need be provided (inlet 60 can communicate with a single outlet 62, which can be larger than the outlets as illustrated) and heating units can be provided. Cooling units can be provided as well, where cooling is desired. Units 20 and 24 can carry any temperature-control fluid, whether to heat or cool.

Referring now to FIG. 5, reaction chamber 16 is shown, including temperature control units 22 and 26, in expanded form. Units 22 and 26 can be the same as units 20 and 24 as shown in FIG. 4, with unit 22 defining the top of reaction chamber 16. Reaction unit 16 includes an inlet 70 fluidly communicating with an outlet 72 and a reaction chamber defined therebetween. The reaction chamber, in microreactor embodiments of the invention, has a volume of less than one milliliter, or other lower volumes as described above in connection with mixing unit 12. Inlet 70 is connectable to a source of a chemical or biological starting material, optionally supplied by mixing unit 12 and heating/dispersion unit 14, and outlet 70 is designed to release the product of a chemical or biological reaction occurring within the chamber involving the starting material. Unit 16 can be formed from materials as described above.

The reactor unit is the core of the process. While the unit is designed to be interchangeable for biological or pharmaceutical reactions, the specific application as shown is for cell cultivation. As in the case of the previous unit, temperature control units such as heat exchanger platforms will sandwich the central reaction chamber. The heat exchangers will maintain the temperature of the reaction unit as the same temperature as discussed for the cell broth.

A feature of the unit is heterogeneous reaction on a supported matrix. Cell feed enters the reaction chamber under the proper pH, O2 concentration, and temperature for cell cultivation. Cells, immobilized onto the silicon framework at locations 74 either by surface functionalization and subsequent reaction or entrapment within a host membrane, metabolize the nutrients provided by the feed stream and produce a product protein. The initial reaction platform can be a two-dimensional array of cells both on the top and bottom of the reaction chamber. This arrangement is to prevent a large pressure drop across the unit which would be detrimental to flow.

In this unit, oxygen and nutrients are diffused from the flowing stream to the immobilized cells. The cells, in turn metabolize the feed, and produce proteins which are swept away in the flowing stream. The flowing stream then enters the fourth chamber which removes the protein product from the solution.

Referring again to FIG. 1, it can be seen how dispersion unit 14 creates an evenly-divided flow of fluid (reactant fluid such as oxygen and nutrients in the case of cell cultivation) across each of locations 74 in reaction to chamber 16.

Referring now to FIG. 6, separation unit 18 is shown in greater detail, in expanded view. Separation unit 18 defines a central unit including an inlet 80 communicating with an outlet 82, and a fluid pathway 84 connecting the inlet with the outlet. Unit 18 can be fabricated as described above with respect to other components of the invention, and preferably is etched silicon. It may be desirable for fluid path 84 to completely span the thickness of unit 18 such that the pathway is exposed both above and below the unit. To maintain structural integrity, pathway 84 can be etched to some extent but not completely through unit 18 as illustrated, and a plurality of holes or channels can be formed through the bottom of the pathway exposing the bottom of the pathway to areas below the unit. Inlet 80 can be connectable to the outlet of reaction chamber 16, and outlet 82 to a container for recovery of carrier fluid.

In the embodiment illustrated, membranes 34 and 36 cover exposed portions of fluid pathway 84 facing upward or downward as illustrated. Membranes 34 and/or 36 can be any membranes suitable for separation, i.e. extraction of product through the membrane with passage of effluent, or carrier fluid, through outlet 82. Those of ordinary skill in the art will recognize a wide variety of suitable membranes including size-selective membranes, ionic membranes, and the like. Upper and lower extraction solvent fluid units 30 and 32, which can comprise materials as described above including etched silicon, each include a fluid pathway 86 connecting an inlet 88 with an outlet 90. Fluid pathway 86 preferably is positioned in register with fluid pathway 84 of unit 18 when the separation unit is assembled. In this way, two flowing streams of solvent through channels 86 of units 30 and 32 flow counter to the direction of flow of fluid in channel 84 of unit 18, the fluids separated only by membranes 34 and 36. This establishes a counter-current tangential flow filtration membrane system. By concentration gradients, products are selectively extracted from channel 84 into solvent streams flowing within channels 86 and unit 30 or 32. Product is recovered through the outlet 90 of units 30 or 32 and recovered in a container (not shown) having a volume that can be greater than 1 liter. Outlets 90 thereby define carrier fluid outlets, and a fluid pathway connects inlet 80 of unit 18 with the carrier fluid outlets 90 of units 30 and 32, breached only by membranes 34 and 36. Carrier fluid outlet 82 can be made connectable to a recovery container for recycling of reaction carrier fluids. In the example of a microbioreactor, residual oxygen and nutrients are recovered from outlet 82 and recycled back into the feed for the process.

The flowing streams of extraction solvent in channels 86 can be set at any desired temperature using temperature control units (not illustrated). In the case of a microbioreactor, these fluids can be set at approximately 4° C. The low temperature is needed to maintain the efficacy of the protein products and prevent denaturation. Additionally, several purification and clarification steps are often performed in industrial application. The necessity of further purification is remedied by the use of additional units in series.

Embedded within the production process can be control systems and detectors for the manipulation of temperature, pH, nutrients, and oxygen concentration. Where a microbiorector is used, the viability of cells is dependent upon strict limits for the parameters mentioned above. Narrow set-point ranges, dependent on the cell system selected, can be maintained using thermocouples, pH detectors, O2 solubility detectors, and glucose detectors between each unit. These measurements will determine the heat exchanger requirements, O2, CO2, N2, and nutrient inputs.

Diaphragm and peristaltic pumps can be used to provide the necessary driving force for fluid flow in the units. Such pumps are also used to maintain flow in the heat exchanger units.

It is a feature of the invention that many of the microreactors as illustrated can be arranged in parallel. Specifically, at least ten reactors can be constructed to operate in parallel, or in other cases at least about 100, 500, 1,000, or even 10,000 reactors can be constructed to operate in parallel. These reactors can be assembled and disassembled as desired.

It is another feature of the invention that individual units 12, 14, 16, and 18 can be constructed and arranged to be connectable to and separable from each other. That is, any arrangement of individual components can be created for a desired reaction. For example, with reference to FIG. 1, heating/dispersion unit 14 may not be necessary. That is, outlet 52 of mixing unit 12 can be connectable to either inlet 60 of heating/dispersion unit 14, or inlet 70 of reaction unit 16 where a heating/dispersion unit is not used. Moreover, assembly and disassembly of reactors to create a system including many, many reactors operating in parallel, as described above, or in series is possible because of the connectability and separability of the components from each other to form systems containing specific desired components, and any number of those or other systems operating together. Equipment for connection and separation of individual components of a reactor can be selected among those known in the art, as can systems for connection of a variety of reactors in parallel or in series. Systems should be selected such that the individual components can be connectable to and separable from each other readily by laboratory or production-facility technicians without irreversible destruction of components such as welding, sawing, or the like. Examples of known systems for making readily reversible connections between components of reactors or between reactors to form parallel reactors or series reactors include male/female interconnections, clips, cartridge housings where components comprise inserts within the housings, screws, or the like.

Those skilled in the art would readily appreciate that all parameters listed herein are meant to be exemplary and that actual parameters will depend upon the specific application for which the methods and apparatus of the present invention are used. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described. In the claims the words “including”, “carrying”, “having”, and the like mean, as “comprising”, including but not limited to.

Patent Citations
Cited PatentFiling datePublication dateApplicantTitle
US4038151Jul 29, 1976Jul 26, 1977Mcdonnell Douglas CorporationCard for use in an automated microbial detection system
US4116775May 3, 1976Sep 26, 1978Mcdonnell Douglas CorporationMachine and process for reading cards containing medical specimens
US4118280May 3, 1976Oct 3, 1978Mcdonnell Douglas CorporationAutomated microbial analyzer
US4318994Feb 26, 1981Mar 9, 1982Mcdonnell Douglas CorporationEnterobacteriaceae species biochemical test card
US4720462Nov 5, 1985Jan 19, 1988Robert RosensonCulture system for the culture of solid tissue masses and method of using the same
US4756884Jul 1, 1986Jul 12, 1988Biotrack, Inc.For detecting presence of analyte in physiological fluid sample
US4839292Sep 11, 1987Jun 13, 1989Cremonese Joseph GCell culture flask utilizing a membrane barrier
US4952373Apr 21, 1989Aug 28, 1990Biotrack, Inc.Liquid shield for cartridge
US5004685Aug 18, 1987Apr 2, 1991Fuji Photo Film Co., Ltd.Dry-type multilayer analytical element
US5047213Jan 18, 1989Sep 10, 1991Amersham International PlcBiological sensors
US5051237Jun 23, 1988Sep 24, 1991P B Diagnostic Systems, Inc.Liquid transport system
US5173225May 7, 1990Dec 22, 1992Bowolfe, Inc.Method for making ultra-thin dense skin membrane
US5219762Dec 20, 1989Jun 15, 1993Mochida Pharmaceutical Co., Ltd.Method and device for measuring a target substance in a liquid sample
US5252294Feb 3, 1992Oct 12, 1993Messerschmitt-Bolkow-Blohm GmbhMicromechanical structure
US5254143Nov 3, 1992Oct 19, 1993Dainippon Ink And Chemical, Inc.Diaphragm for gas-liquid contact, gas-liquid contact apparatus and process for producing liquid containing gas dissolved therein
US5278048May 29, 1991Jan 11, 1994Molecular Devices CorporationMeasuring a change in PH
US5387329Apr 9, 1993Feb 7, 1995Ciba Corning Diagnostics Corp.Extended use planar sensors
US5424209Mar 19, 1993Jun 13, 1995Kearney; George P.Noncontamination; controlled supply of nutrients; supplying life sustaining gases; venting waste gases; pressurization; heating and cooling control
US5430542Apr 10, 1992Jul 4, 1995Avox Systems, Inc.Disposable optical cuvette
US5436129Oct 12, 1993Jul 25, 1995Gene Tec Corp.Automated detection of DNA sequences
US5449617Sep 2, 1993Sep 12, 1995Heraeus Sepatech GmbhCulture vessel for cell cultures
US5478751Apr 18, 1994Dec 26, 1995Abbott LaboratoriesDetermining presence or amount of analyte from detectable signal
US5496697Sep 8, 1993Mar 5, 1996Molecular Devices CorporationMethods and apparatus for detecting the effect of cell affecting agents on living cells
US5534328Dec 2, 1993Jul 9, 1996E. I. Du Pont De Nemours And CompanyIntegrated chemical processing apparatus and processes for the preparation thereof
US5576211Apr 18, 1995Nov 19, 1996Heraeus Instruments GmbhNutrients for culture
US5580523Apr 1, 1994Dec 3, 1996Bard; Allen J.Integrated chemical synthesizers
US5587128Nov 14, 1994Dec 24, 1996The Trustees Of The University Of PennsylvaniaPolymerase chain reaction
US5589350Jan 11, 1996Dec 31, 1996Biolog, Inc.Testing device for liquid and liquid suspended samples
US5593632May 18, 1995Jan 14, 1997Seiji KagawaMethod of making a porous film
US5595712Jul 25, 1994Jan 21, 1997E. I. Du Pont De Nemours And CompanyChemical mixing and reaction apparatus
US5602028Jun 30, 1995Feb 11, 1997The University Of British ColumbiaUsed to assay drugs for their activity and ability to penetrate tissue
US5612188Feb 10, 1994Mar 18, 1997Cornell Research Foundation, Inc.Automated, multicompartmental cell culture system
US5632957Sep 9, 1994May 27, 1997NanogenMolecular biological diagnostic systems including electrodes
US5639423Aug 31, 1992Jun 17, 1997The Regents Of The University Of CalforniaChamber with thin film wall with element on wall for manipulation of parameter of chemical reaction
US5646039Jun 6, 1995Jul 8, 1997The Regents Of The University Of CaliforniaBiochemical reactions
US5674742Jun 6, 1995Oct 7, 1997The Regents Of The University Of CaliforniaMicrofabricated reactor
US5705018Dec 13, 1995Jan 6, 1998Hartley; Frank T.Micromachined peristaltic pump
US5744366Nov 14, 1994Apr 28, 1998Trustees Of The University Of PennsylvaniaMesoscale devices and methods for analysis of motile cells
US5800785Jul 24, 1996Sep 1, 1998Biolog, Inc.Testing device for liquid and liquid suspended samples
US5800788Oct 3, 1995Sep 1, 1998Sandvik AbReactor container, plant and process for the production of sulfuric acid
US5840258Jul 12, 1993Nov 24, 1998Foster Wheeler Energia OyMethod and apparatus for transporting solid particles from one chamber to another chamber
US5842787Oct 9, 1997Dec 1, 1998Caliper Technologies CorporationMicrofluidic systems incorporating varied channel dimensions
US5846396Nov 9, 1995Dec 8, 1998Sarnoff CorporationLiquid distribution system
US5856174Jan 19, 1996Jan 5, 1999Affymetrix, Inc.Integrated nucleic acid diagnostic device
US5858770Sep 30, 1997Jan 12, 1999Brandeis UniversityCell culture plate with oxygen and carbon dioxide-permeable waterproof sealing membrane
US5916812May 13, 1997Jun 29, 1999Biomerieux, Inc.Test sample card with polymethylpentene tape
US5928880Jun 11, 1997Jul 27, 1999Trustees Of The University Of PennsylvaniaSeparating target cells from a fluid sample using bound proteins in multicompartmented container; quick, efficient diagnosis
US5942443Jun 28, 1996Aug 24, 1999Caliper Technologies CorporationUsing substrate having at least two intersecting channels, continuously flowing biochemical system through one channel, flowing test compound from second channel into first, detecting effect of test compound on system
US5957579Sep 30, 1998Sep 28, 1999Caliper Technologies Corp.Microfluidic systems incorporating varied channel dimensions
US5958694Oct 16, 1997Sep 28, 1999Caliper Technologies Corp.Microscale separation channel having first and second ends for separating nucleic acid fragments by size; nested sets of first and second nucleotide termination fragments at different concentrations connected to separation channel
US5964995Apr 4, 1997Oct 12, 1999Caliper Technologies Corp.Enhancing material direction and transport by electroosmotic flow of a fluid containing that material by adding a zwitterionic compound; reducing electrophoretic separation of differentially charged species in a microscale fluid column
US5965092Oct 15, 1997Oct 12, 1999Eastman Kodak CompanyIntegrated micro-ceramic chemical plant with insertable micro-filters
US5972187Mar 20, 1998Oct 26, 1999Caliper Technologies CorporationIntersecting capillary channels arranged so that a sample fluid being transported along the first channel towards the second channel is mixed at intersection and bias in the fluid is dissipated
US5976472Oct 15, 1997Nov 2, 1999Eastman Kodak CompanyIntegrated micro-ceramic chemical plant with insertable catalytic reaction chambers
US5981211Oct 7, 1996Nov 9, 1999Regents Of The University Of MinnesotaContinuous production of cell product from animal tissues or geneticaly engineered cells by trapping cells in insoluble, cell compatible matrix; supplying nutrient medium; withdrawing waste; withdrawing prodduct
US5985119May 10, 1996Nov 16, 1999Sarnoff CorporationElectrokinetic pumping
US5989835Feb 27, 1997Nov 23, 1999Cellomics, Inc.Automated monitoring of fluorescence; analyzing using computer; data processing
US5992769Jun 9, 1995Nov 30, 1999The Regents Of The University Of MichiganMicrochannel system for fluid delivery
US5993750Apr 11, 1997Nov 30, 1999Eastman Kodak CompanyIntegrated ceramic micro-chemical plant
US6001352Mar 31, 1997Dec 14, 1999Osteobiologics, Inc.Contacting condrocytes with specific platelet-derived growth factors (pdgf) in the absence of growth factors which promote cell differentiation; cells cultured and loaded onto scaffolding for implanting into a cartilage or bone wound
US6001585Nov 14, 1997Dec 14, 1999Cellex Biosciences, Inc.Biological apparatus made of oxygen permeable tubing that has interior and exterior spacing and lacks a pump; for efficient screening and propagation of cells
US6008010Nov 1, 1996Dec 28, 1999University Of PittsburghA cell culturing apparatus; incubating cells in biochamber that automatically controls the closed environment, cells are maintained at desired conditions and can be examined while culturing; automated growth of stem cells; cancer therapy
US6012902Sep 25, 1997Jan 11, 2000Caliper Technologies Corp.Micropump
US6025601Jun 9, 1997Feb 15, 2000Affymetrix, Inc.Method and apparatus for imaging a sample on a device
US6042710May 11, 1999Mar 28, 2000Caliper Technologies Corp.Methods and compositions for performing molecular separations
US6043080Dec 11, 1998Mar 28, 2000Affymetrix, Inc.One piece, multicompartment device having a chamber for amplification, one for fragmentation, and another with array of probes coupled to substrate
US6046056Dec 6, 1996Apr 4, 2000Caliper Technologies CorporationHigh throughput screening assay systems in microscale fluidic devices
US6090251Jun 6, 1997Jul 18, 2000Caliper Technologies, Inc.Microfabricated structures for facilitating fluid introduction into microfluidic devices
US6103199Sep 15, 1998Aug 15, 2000Aclara Biosciences, Inc.Capillary electroflow apparatus and method
US6103479May 29, 1997Aug 15, 2000Cellomics, Inc.Miniaturized cell array methods and apparatus for cell-based screening
US6107044Jun 16, 1999Aug 22, 2000Caliper Technologies Corp.Apparatus and methods for sequencing nucleic acids in microfluidic systems
US6117643Nov 25, 1997Sep 12, 2000Ut Battelle, LlcBioluminescent bioreporter integrated circuit
US6126946Jul 23, 1997Oct 3, 2000University Of Michigan, The Board Of RegentsHepatocyte-selective oil-in-water emulsion
US6143247Dec 19, 1997Nov 7, 2000Gamera Bioscience Inc.Affinity binding-based system for detecting particulates in a fluid
US6148968Jun 1, 1999Nov 21, 2000Dana CorporationIntegrated axle spindle and brake spider
US6150180Jul 26, 1999Nov 21, 2000Caliper Technologies Corp.High throughput screening assay systems in microscale fluidic devices
US6167910Jan 14, 1999Jan 2, 2001Caliper Technologies Corp.Multi-layer microfluidic devices
US6171067Oct 20, 1999Jan 9, 2001Caliper Technologies Corp.Micropump
US6171850Mar 8, 1999Jan 9, 2001Caliper Technologies Corp.Apparatus for accurate analysis of reactions in a heat controlled environment
US6174675Aug 27, 1998Jan 16, 2001Caliper Technologies Corp.Multicompartment apparatus for monitoring and controlling processes parameters; for use as diagnostic tools in genetic engineering
US6176962Jun 18, 1997Jan 23, 2001Aclara Biosciences, Inc.Methods for fabricating enclosed microchannel structures
US6184029Jan 27, 1999Feb 6, 2001Trustees Of The University Of PennsylvaniaApparatus for the detection and analysis of particles or microorganisms in sample
US6186660Jul 26, 1999Feb 13, 2001Caliper Technologies Corp.Microfluidic systems incorporating varied channel dimensions
US6193647Apr 8, 1999Feb 27, 2001The Board Of Trustees Of The University Of IllinoisMicrofluidic embryo and/or oocyte handling device and method
US6221226Oct 7, 1999Apr 24, 2001Caliper Technologies Corp.Methods and systems for monitoring and controlling fluid flow rates in microfluidic systems
US6221654Sep 23, 1997Apr 24, 2001California Institute Of TechnologyChip of substrate and analysis unit; unit has two branch channels, detector, flow controller, and main channel with sample inlet, detection region for one polynucleotide at a time, and branch point discrimination region
US6235175Oct 2, 1998May 22, 2001Caliper Technologies Corp.Microfluidic devices incorporating improved channel geometries
US6238538Apr 6, 1999May 29, 2001Caliper Technologies, Corp.Controlled fluid transport in microfabricated polymeric substrates
US6245295Feb 23, 1999Jun 12, 2001Bio Merieux, Inc.Sample testing apparatus with high oxygen permeable tape providing barrier for closed reaction chamber
US6251343Feb 24, 1998Jun 26, 2001Caliper Technologies Corp.Microfluidic devices and systems incorporating cover layers
US6264892Jan 11, 2000Jul 24, 2001Agilent Technologies, Inc.Miniaturized planar columns for use in a liquid phase separation apparatus
US6267858Jun 24, 1997Jul 31, 2001Caliper Technologies Corp.High throughput screening assay systems in microscale fluidic devices
US6274089Jun 8, 1998Aug 14, 2001Caliper Technologies Corp.Microfluidic channel network in substrate surface having a reaction region and a separation region whereby the reagent flow is controlled by electrokinetics
US6274337Mar 19, 1998Aug 14, 2001Caliper Technologies Corp.High throughput screening assay systems in microscale fluidic devices
US6319469Dec 18, 1996Nov 20, 2001Silicon Valley BankMicrosystem; platform, substrate with embedded reaction chambers and channels connected to microvalve and vented thereby, fluid flow controlled; micromanipulation device interface; microanalyses and microsynthesis
US6319474Jun 7, 1999Nov 20, 2001The Regents Of The University Of CaliforniaMicrofabricated instrument for tissue biopsy and analysis
US6321791Oct 4, 2000Nov 27, 2001Caliper Technologies Corp.Multi-layer microfluidic devices
US6338790Apr 21, 1999Jan 15, 2002Therasense, Inc.Small volume in vitro analyte sensor with diffusible or non-leachable redox mediator
US6339023Apr 6, 1999Jan 15, 2002Applied Materials Inc.Multistep cvd from a mixed gas containing tungsten hexafluoride and silane; in third step adding hydrogen to gas mix
US6551841 *Jan 27, 1999Apr 22, 2003The Trustees Of The University Of PennsylvaniaSmall, typically single-use, modules capable of receiving and rapidly conducting a predetermined assay protocol on a fluid sample
Non-Patent Citations
Reference
1Hayakawa, Y., et al. "Synthesis of Poly (phenylacetylene)s Containing Trifluoromethyl Groups for Gas Permeable Membrane," Journal of Polymer Science, vol. 30, pp. 873-877.
2Hayakawa, Y., et. al, "Synthesis of Poly(phenylacetylene)s Containing Trifluoromethyl Group for Gas Permeable Membrane," Journal of Polymer Science: Part A: Polymer Chemistry, vol. 30, pp. 873-877 (1992).
3Hediger, S., et al., "Biosystem for the culture and characterisation of epithelial cell tissues," Sensors and Actuators B 63 (2000) pp. 63-73.
4International Preliminary Examination Report for International Application Serial No. PCT/US01/07679, published as International Publication No. WO 01/68257, dated May 20, 2002.
5International Preliminary Examination Report for International Application Serial No. PCT/US02/11422, published as International Publication No. WO 02/083852, dated Nov. 21, 2003.
6International Search Report dated Dec. 2, 2005, for PCT/US05/019914, filed Jun. 7, 2005.
7International Search Report dated Jan. 12, 2004 in International Application No. PCT/US03/17816.
8International Search Report for International Application Serial No. PCT/US01/07679, published as International Publication No. WO 01/68257, dated Jun. 8, 2001.
9International Search Report for International Application Serial No. PCT/US02/11422, published as International Publication No. WO 02/083852, dated Mar. 12, 2003.
10Office Action dated Apr. 17, 2007 in U.S. Appl. No. 10/664,046.
11Office Action dated Feb. 7, 2006 from U.S. Appl. No. 10/119,917.
12Office Action dated Jan. 11, 2005 from U.S. Appl. No. 10/119,917.
13Office Action dated Jul. 24, 2007 in U.S. Appl. No. 10/664,067.
14Office Action dated May 19, 2004 from U.S. Appl. No. 10/119,917.
15Office Action dated Oct. 6, 2003 from U.S. Appl. No. 10/119,917.
16Pinnau, I., et al. "Influence of Side-Chain Length on the Gas Permeation Properties of Poly(2-alkylacetylenes)," Macromolecules, vol. 37, pp. 2823-2828.
17Pinnau, I., et al., "Influence of Side-Chain Length on the Gas Permeation Properties of Poly(2-alkylacetylenes)," Macromolecules, vol. 37, pp. 2823-2828 (2004).
18Searby et al., "Space Life Support From The Cellular Perspective," ICES Proceedings, May 2001.
19Written Opinion for International Application Serial No. PCT/US01/07679, published as International Publication No. WO 01/68257, dated Jan. 11, 2002.
20Written Opinion for International Application Serial No. PCT/US02/11422, published as International Publication No. WO 02/083852, dated May 5, 2003.
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US8517596Mar 6, 2012Aug 27, 2013International Business Machines CorporationUsing a microfluid mixer
US8585280Jan 28, 2013Nov 19, 2013International Business Machines CorporationManufacturing a microfluid mixer
US20100003745 *Jun 29, 2009Jan 7, 2010Canon Kabushiki KaishaCell culture vessel
US20110130310 *Mar 31, 2009Jun 2, 2011Technische Universitat IlmenauMicrobioreactor and microtiter plate comprising a plurality of microbioreactors
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Classifications
U.S. Classification435/288.5, 435/297.1, 435/294.1, 435/286.7, 435/297.5
International ClassificationB81B1/00, C12P21/00, B01F13/00, C12P1/00, B01F5/06, G01N37/00, C12M3/00, G01N33/53, B01F5/02, B81B7/04, B01J19/00, B01J19/24, C12M1/00, C12M1/12
Cooperative ClassificationB01J2219/00862, B01F2005/0621, B01J2219/00963, B01F5/0256, B01J2219/00871, B01F13/0064, B01J2219/00889, B01J2219/00961, B01J2219/00966, B01F15/00935, B01J2219/00873, B01J2219/00907, B01F13/0094, B01F5/0646, B01J2219/00957, B01F5/0647, B01F5/061, B01J2219/00835, C12M23/16, B01J2219/00783, B01J2219/00828, B01J19/0093
European ClassificationB01F13/00M6J, B01F15/00T2, B01F5/06B3F, C12M23/16, B01F5/06B3F2, B01J19/00R, B01F5/06B3B, B01F13/00M2B, B01F5/02C
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Effective date: 20001210